Service Principle

The main principle of adenovirus packaging technology is to clone the target fragment into the adenovirus expression vector, the expression vector and packaging plasmid are recombined into adenovirus packaging cells for packaging, and the packaged virus is further amplified and titered.

Service Introduction

Adenovirus is a linear double-stranded DNA virus. Due to its wide host range, ability to infect dividing and non-dividing cells, high titer preparation, and large loading capacity of foreign genes, recombinant adenovirus has been more and more widely used. It is used in basic research and clinical trials, and has become one of the most commonly used gene therapy vectors in the world.

Service Advantage

·There are many types of cells that can be infected and a wide range of hosts;

·High infection efficiency, especially suitable for cells with low transfection efficiency of plasmid vectors, such as primary cells, nerve cells, etc., and the expression level of the target protein is high;

·Can package large fragments of foreign genes, up to 10kb;

·Not integrated into cell chromosomes, high biological safety.

Reqbio has rich experience in virus packaging and has successfully packaged the required viruses for many customers. The company provides two more commonly used systems: AdEasy and AdMAX.

AdEasy system:

The main feature of the AdEasy packaging system is to use the Cre/loxP system to complete the process of inserting foreign genes into the adenovirus genome in Escherichia coli (BJ5183), obtain a circular recombinant adenovirus genome, and have the necessary elements for replication in bacteria. The recombinant adenovirus genome was digested and linearized and then transfected into HEK293 cells, avoiding dual plasmid co-transfection to obtain recombinant adenovirus.

The following issues need to be paid attention to when using this system:

1. To use specific Escherichia coli that can express Cre recombinase, it is recommended to establish a strain bank to avoid bacterial changes;

2. Since recombination occurs in bacteria, small genome changes are not easy to detect;

3. To obtain high-quality recombinant adenovirus seeds (high virus yield and high target gene expression), multiple recombinant adenovirus plasmids should be selected and transfected into HEK293 cells to obtain multiple recombinant viruses for screening.

Adenoviral shuttle vectors can be constructed according to different research needs, such as adenoviral vectors for RNAi, adenoviral vectors for gene overexpression, adenoviral vectors for promoter activity research, etc.

AdMAX system:

The AdMax packaging system obtains recombinant virus through Cre/loxP, which occurs in HEK293 cells, thereby avoiding recombination in bacteria. The adenovirus shuttle plasmid cloned with foreign genes and the packaging plasmid carrying most of the adenovirus genome were co-transfected into HEK293 cells, and the Cre/loxP system was used to achieve recombination to produce recombinant adenovirus. This system has many advantages, including easy operation, high recombination efficiency, high yield of virus obtained, and high expression level of the target gene (because the mCMV promoter is several times stronger than the hCMV promoter), etc.

Adenoviral shuttle vectors can be constructed according to different research needs, such as adenoviral vectors for RNAi, adenoviral vectors for gene overexpression, adenoviral vectors for promoter activity research, etc.

Service flow

1) Obtain the target gene sequence;

2) Construction of viral expression vector plasmids;

3) Expression vector plasmid and packaging plasmid recombination;

4) Infect HEK293 and package the virus;

5) Virus amplification and concentration;

6) Titer determination.

Service Description

1) The customer provides the sequence of the target gene, and if a template is provided, please inform the carrier, enzyme digestion stability point and other information;

2) The AdEasy system is used by default, please let us know if you have specific requirements;

3) The sequence is too long may affect the virus titer;

4) The functional verification of the target protein will be charged additionally.

Ordering method

  Service Hotline：4008-750-250                                  Phone：180-6607-1954

  QQ：4008-750-250                                                     Mail：sales@reqbio.com